Although Dr. Mario does not appear in the World of Light opening cutscene, he was vaporized and later imprisoned alongside the rest of the fighters (sans Kirby) when Galeem unleashed his beams of light. Dr. Mario has his own awakening battle separate from that of Mario, who appears in the opening cutscene.

Dr. Mario can be awakened in a cloud that is found on a pathway connected by a ribbon that also leads to a city-like area.